SOP - Stanford Behavioral and Functional Neuroscience Laboratory

Stanford Behavioral and Functional
Neuroscience Laboratory
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STANDARD OPERATING PROCEDURE
TITLE: Garcia Neurological Test
CATEGORY: Behavioral Assay
Introduction
Goal: This document aims to provide the reader information on how to conduct the Garcia
Neurological Test. The test can be run simultaneously with the 28-point Neuroscore Test.
Scores for the following six tests are summed to give a maximum total score of 18 points:
1. Spontaneous activity (max score 3)
2. Symmetry in forelimb movement (max score 3)
3. Forepaw outstretching (max score 3)
4. Climbing (max score 3)
5. Body proprioception (max score 3)
6. Response to vibrissae touch (max score 3)
As this is a general description of standard materials, test settings, and procedures, variations
may be made to fit specific needs.
Materials
 Subjects: any strain of rats. No prior training is required, though subjects should be
acclimated to testing environment and experimenter before testing.
 Wire sheet: used in climbing test.
 Stick: used in body proprioception and response to vibrissae touch tests.
 10% ethanol: used to clean between trials.
Test Settings
 Test area: clean table top with sufficient space for animal and two home cages. No
additional people in the room during testing.
 Lighting: well-lit.
TITLE: Garcia Neurological Test
Stanford Behavioral and Functional
Neuroscience Laboratory
CATEGORY: Behavioral Assay
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Version 4.0
Detailed Standard Operating Procedure
Before testing:
 Acclimation: subjects in home cage are placed in testing room for at least 1hr before
testing to minimize effects of stress on behavior during testing.
 Subject training: none required.
Testing procedures:
 Animal is subjected to a number of tests, with sequence of testing unimportant.
Behavior is scored for each test according to score systems presented in Appendix A.
 Animal is returned to home cage and test area is cleaned with 10% ethanol between
trials.
Data Analysis
 Scores for each test are summed to determine the overall Garcia test score (possible
scores range from 3 to 18).
Appendix A: Scoring
Each score system may be broken down into half-point scores.
1. Spontaneous activity
(Activity is observed in test home cage for 5min.)
3
Normal: approaches all four walls of cage
2
Slightly affected: hesitates but still explores walls
1
Moderately affected: barely moves but eventually approaches wall
0
Severely affected: does not move
2. Symmetry in the movement of forepaws
(Outstretching and movement of forepaws are observed when animal is lifted by base of tail
above ground.)
3
Normal: forepaws are symmetrical
2
Slight: limbs on impaired side do not extend as much as on normal side
1
Moderate: limbs on impaired side do not extend but are held close to body
0
Severe: limbs on impaired side do not move at all
3. Forepaw outstretching
(Forepaw outstretching and forepaw walking are observed when animal is lifted by base of tail
and allowed to walk on forepaws across table. Forepaw use may also be observed when animal
is dragged backwards by base of tail.)
TITLE: Garcia Neurological Test
Stanford Behavioral and Functional
Neuroscience Laboratory
CATEGORY: Behavioral Assay
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Normal: forepaws are outstretched and animal is able to walk on forepaws in
straight line
Slight: forepaws are outstretched and animal walks on forepaws but towards
impaired side
Moderate: impaired forepaw does not outstretch well and forepaw walking is
heavily biased toward impaired side
Severe: impaired forepaw does not outstretch or move during walking
4. Climbing
(Climbing is observed when animal is lifted by base of tail and placed on wire sheet resting on
side of home cage.)
3
Normal: grips wire sheet and climbs up to home cage with ease
2
Moderate: climbs up to home cage but impaired paw misses wires while climbing
1
Severe: cannot grip wire sheet with impaired paw and cannot climb up to home
cage
5. Body proprioception
(Reaction is observed when each side of animal is poked separately while crawling on floor.
Animal should not see stick before poking.)
3
Normal: inspects both sides or is startled after poking on either side
2
Moderate: inspects impaired side after poking more slowly than unaffected side
1
Severe: does not respond after being poked on impaired side
6. Response to vibrissae touch
(Reaction is observed when whiskers are brushed on each side separately with a long, thin stick.
Stick should approach from the rear and animal should be distracted with hand in front of head
to ensure animal does not see stick while testing. Behavior should be scored during first few
brushes, as too many brushes will accustom the animal to whiskers being touched.)
3
Normal: brushes side of head after whiskers are touched with stick, or head
retreats from stick
2
Moderate: head retreats from stick more slowly after brushing whiskers on
affected side as compared to unaffected side
1
Severe: does not respond after brushing whiskers on impaired side
TITLE: Garcia Neurological Test
Stanford Behavioral and Functional
Neuroscience Laboratory
CATEGORY: Behavioral Assay
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Version 4.0
Example Garcia Test Score Sheet
Study #:
Garcia Test
Group:
Test #:
Animal
ID#
Forepaw
Sym.
Forepaw
Stretch
Spont.
Activity
Climbing
Date:
Initials:
Body
Touch
Vibrissae
Touch
Sum

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